Triazabenzo[E]Azulene Derivatives for the Treatment Of Tumors

ABSTRACT

Novel triazabenzo[e]azulene derivatives of the formula, (I) in which R 1 , R 2  and R 3  have the meanings indicated in Claim  1,  are inhibitors of TGF-beta receptor kinase and can be employed, inter alia, for the treatment of tumours.

BACKGROUND OF THE INVENTION

The invention had the object of finding novel compounds having valuableproperties, in particular those which can be used for the preparation ofmedicaments.

The present invention relates to compounds and to the use of compoundsin which the inhibition, regulation and/or modulation of signaltransduction by kinases, in particular TGF-beta receptor kinases, playsa role, furthermore to pharmaceutical compositions which comprise thesecompounds, and to the use of the compounds for the treatment ofkinase-induced diseases.

Transforming growth factor beta is the prototype of the TGF-betasuperfamily, a family of highly preserved, pleiotropic growth factors,which carry out important functions both during embryo development andalso in the adult organism. In mammals, three isoforms of TGF-beta(TGF-beta 1, 2 and 3) have been identified, TGF-beta 1 being thecommonest isoform (Kingsley (1994) Genes Dev 8:133-146). TGF-beta 3 isexpressed, for example, only in mesenchymal cells, whereas TGF-beta 1 isfound in mesenchymal and epithelial cells. TGF-beta is synthesised aspre-proprotein and is released in inactive form into the extracellularmatrix (Derynck (1985) Nature 316: 701-705; Bottinger (1996) PNAS 93:5877-5882). Besides the proregion cleaved off, which is also known aslatency associated peptide (LAP) and remains associated with the matureregion, one of the 4 isoforms of the latent TGF-beta binding proteins(LTBP 1-4) may also be bonded to TGF-beta (Gentry (1988) Mol Cell Biol8: 4162-4168, Munger (1997) Kindey Int 51: 1376-1382). The activation ofthe inactive complex that is necessary for the development of thebiological action of TGF-beta has not yet been clarified in full.However, proteolytic processing, for example by plasmin, plasmatransglutaminase or thrombospondin, is certainly necessary (Munger(1997) Kindey Int 51: 1376-1382). The activated ligand TGF-beta mediatesits biological action via three TGF-beta receptors on the membrane, theubiquitously expressed type I and type II receptors and the type IIIreceptors betaglycan and endoglin, the latter only being expressed inendothelial cells (Gougos (1990) J Biol Chem 264: 8361-8364,Loeps-Casillas (1994) J Cell Biol 124:557-568). Both type III TGF-betareceptors lack an intracellular kinase domain which facilitates signaltransmission into the cell. Since the type III TGF-beta receptors bindall three TGF-beta isoforms with high affinity and type II TGF-betareceptor also has higher affinity for ligands bonded to type IIIreceptor, the biological function is thought to consist in regulation ofthe availability of the ligands for type I and type II TGF-betareceptors (Lastres (1996) J Cell Biol 133:1109-1121; Lopes-Casillas(1993) Cell 73: 1435-1344). The structurally closely related type I andtype II receptors have a serine/threonine kinase domain, which isresponsible for signal transmission, in the cytoplasmatic region. TypeII TGF-beta receptor binds TGF-beta, after which the type I TGF-betareceptor is recruited to this signal-transmitting complex. Theserine/threonine kinase domain of the type II receptor is constitutivelyactive and is able to phosphorylate seryl radicals in this complex inthe so-called GS domain of the type I receptor. This phosphorylationactivates the kinase of the type I receptor, which is now itself able tophosphorylate intracellular signal mediators, the SMAD proteins, andthus initiates intracellular signal transmission (summarised in Derynck(1997) Biochim Biophys Acta 1333: F105-F150).

The proteins of the SMAD family serve as substrates for all TGF-betafamily receptor kinases. To date, 8 SMAD proteins have been identified,which are divided into 3 groups: (1) receptor-associated SMADs (R-SMADs)are direct substrates of the TGF-β receptor kinases (SMAD1, 2, 3, 5, 8);(2) co-SMADs, which associate with the R-Smads during the signal cascade(SMAD4); and (3) inhibitory SMADs (SMAD6, 7), which inhibit the activityof the above-mentioned SMAD proteins. Of the various R-SMADs, SMAD2 andSMAD3 are the TGF-beta-specific signal mediators. In the TGF-beta signalcascade, SMAD2/SMAD3 are thus phosphorylated by the type I TGF-betareceptor, enabling them to associate with SMAD4. The resultant complexof SMAD2/SMAD3 and SMAD4 can now be translocated into the cell nucleus,where it can initiate the transcription of the TGF-beta-regulated genesdirectly or via other proteins (summarised in Itoh (2000) Eur J Biochem267: 6954-6967; Shi (2003) Cell 113: 685-700).

The spectrum of the functions of TGF-beta is wide-ranging and dependenton cell type and differentiation status (Roberts (1990) Handbook ofExperimental Pharmacology: 419-472). The cellular functions which areinfluenced by TGF-beta include: apoptosis, proliferation,differentiation, mobility and cell adhesion. Accordingly, TGF-beta playsan important role in a very wide variety of biological processes. Duringembryo development, it is expressed at sites of morphogenesis and inparticular in areas with epithelial-mesenchymal interaction, where itinduces important differentiation processes (Pelton (1991) J Cell Biol115:1091-1105). TGF-beta also carries out a key function in theself-renewal and maintenance of an undifferentiated state of stem cells(Mishra (2005) Science 310: 68-71). In addition, TGF-beta also fulfilsimportant functions in the regulation of the immune system. It generallyhas an immunosuppressive action, since it inhibits, inter alia, theproliferation of lymphocytes and restricts the activity of tissuemacrophages. TGF-beta thus allows inflammatory reactions to subsideagain and thus helps to prevent excessive immune reactions (Bogdan(1993) Ann NY Acad Sci 685: 713-739, summarised in Letterio (1998) AnnuRev Immunol 16: 137-161). Another function of TGF-beta is regulation ofcell proliferation. TGF-beta inhibits the growth of cells ofendothelial, epithelial and haematopoietic origin, but promotes thegrowth of cells of mesenchymal origin (Tucker (1984) Science226:705-707, Shipley (1986) Cancer Res 46:2068-2071, Shipley (1985) PNAS82: 4147-4151). A further important function of TGF-beta is regulationof cellular adhesion and cell-cell interactions. TGF-beta promotes thebuild-up of the extracellular matrix by induction of proteins of theextracellular matrix, such as, for example, fibronectin and collagen. Inaddition, TGF-beta reduces the expression of matrix-degradingmetalloproteases and inhibitors of metalloproteases (Roberts (1990) AnnNY Acad Sci 580: 225-232; Ignotz (1986) J Biol Chem 261: 4337-4345;Overall (1989) J Biol Chem 264: 1860-1869); Edwards (1987) EMBO J 6:1899-1904).

The broad spectrum of action of TGF-beta implies that TGF-beta plays animportant role in many physiological situations, such as wound healing,and in pathological processes, such as cancer and fibrosis.

TGF-beta is one of the key growth factors in wound healing (summarisedin O'Kane (1997) Int J Biochem Cell Biol 29: 79-89). During thegranulation phase, TGF-beta is released from blood platelets at the siteof injury. TGF-beta then regulates its own production in macrophages andinduces the secretion of other growth factors, for example by monocytes.The most important functions during wound healing include stimulation ofchemotaxis of inflammatory cells, the synthesis of extracellular matrixand regulation of the proliferation, differentiation and gene expressionof all important cell types involved in the wound-healing process.

Under pathological conditions, these TGF-beta-mediated effects, inparticular the regulation of the production of extracellular matrix(ECM), can result in fibrosis or scars in the skin (Border (1994) N EnglJ Med 331:1286-1292).

For the fibrotic diseases, diabetic nephropathy and glomeronephritis, ithas been shown that TGF-beta promotes renal cell hypertrophy andpathogenic accumulation of the extracellular matrix. Interruption of theTGF-beta signalling pathway by treatment with anti-TGF-beta antibodiesprevents expansion of the mesangial matrix, progressive reduction inkidney function and reduces established lesions of diabeticglomerulopathy in diabetic animals (Border (1990) 346: 371-374, Yu(2004) Kindney Int 66: 1774-1784, Fukasawah (2004) Kindney Int 65:63-74, Sharma (1996) Diabetes 45: 522-530).

TGF-beta also plays an important role in liver fibrosis. The activation,essential for the development of liver fibrosis, of the hepatic stellatecells to give myofibroblasts, the main producer of the extracellularmatrix in the course of the development of liver cirrhosis, isstimulated by TGF-beta. It has likewise been shown here thatinterruption of the TGF-beta signalling pathway reduces fibrosis inexperimental models (Yata (2002) Hepatology 35:1022-1030; Arias (2003)BMC Gastroenterol 3:29)

TGF-beta also takes on a key function in the formation of cancer(summarised in Derynck (2001) Nature Genetics: 29: 117-129; Elliott(2005) J Clin Onc 23: 2078-2093). In early stages of the development ofcancer, TGF-beta counters the formation of cancer. Thistumour-suppressive action is based principally on the ability ofTGF-beta to inhibit the division of epithelial cells. By contrast,TGF-beta promotes cancer growth and the formation of metastases in latetumour stages. This can be attributed to the fact that most epithelialtumours develop a resistance to the growth-inhibiting action ofTGF-beta, and TGF-beta simultaneously supports the growth of the cancercells via other mechanisms. These mechanisms include promotion ofangiogenesis, the immunosuppressive action, which supports tumour cellsin avoiding the control function of the immune system(immunosurveillance), and promotion of invasiveness and the formation ofmetastases. The formation of an invasive phenotype of the tumour cellsis a principal prerequisite for the formation of metastases. TGF-betapromotes this process through its ability to regulate cellular adhesion,motility and the formation of the extracellular matrix. Furthermore,TGF-beta induces the transition from an epithelial phenotype of the cellto the invasive mesenchymal pheno-type (epithelial mesenchymaltransition=EMT). The important role played by TGF-beta in the promotionof cancer growth is also demonstrated by investigations which show acorrelation between strong TGF-beta expression and a poor prognosis.Increased TGF-beta level have been found, inter alia, in patients withprostate, breast, intestinal and lung cancer (Wikström (1998) Prostate37:19-29; Hasegawa (2001) Cancer 91: 964-971; Friedman (1995), CancerEpidemiol Biomarkers Prey. 4:549-54).

Owing to the cancer-promoting actions of TGF-beta described above,inhibition of the TGF-beta signalling pathway, for example viainhibition of the TGF-beta type I receptor, is a possible therapeuticconcept. It has been shown in numerous preclinical trials thatinterruption of the TGF-beta signalling pathway does indeed inhibitcancer growth. Thus, treatment with soluble TGF-beta type II receptorreduces the formation of metastases in transgenic mice, which developinvasive breast cancer in the course of time (Muraoka (2002) J ClinInvest 109: 1551-1559, Yang (2002) J Clin Invest 109: 1607-1615).

Tumour cell lines which express a defective TGF-beta type II receptorexhibit reduced tumour and metastatic growth (Oft (1998) Curr Biol 8:1243-1252, McEachern (2001) Int J Cancer 91:76-82, Yin (1999) JclinInvest 103: 197-206).

Conditions “characterised by increased TGF-β activity” include those inwhich TGF-β synthesis is stimulated so that TGF-β is present atincreased levels or in which latent TGF-β protein is undesirablyactivated or converted to active TGF-β protein or in which TGF-βreceptors are upregulated or in which the TGF-β protein shows enhancedbinding to cells or the extracellular matrix in the location of thedisease. Thus, in each case “increased activity” refers to any conditionin which the biological activity of TGF-β is undesirably high,regardless of the cause.

A number of diseases have been associated with TGF-β1 overproduction.Inhibitors of the intracellular TGF-β signalling pathway are suitabletreatments for fibroproliferative diseases. Specifically,fibroproliferative diseases include kidney disorders associated withunregulated TGF-β activity and excessive fibrosis includingglomerulonephritis (GN), such as mesangial proliferative GN, immune GNand crescentic GN. Other renal conditions include diabetic nephropathy,renal interstitial fibrosis, renal fibrosis in transplant patientsreceiving cyclosporin, and HIV-associated nephropathy. Collagen vasculardisorders include progressive systemic sclerosis, polymyositis,sclerodermatitis, dermatomyositis, eosinophilic fasciitis, morphea, orthose associated with the occurrence of Raynaud's syndrome. Lungfibroses resulting from excessive TGF-β activity include adultrespiratory distress syndrome, idiopathic pulmonary fibrosis, andinterstitial pulmonary fibrosis often associated with autoimmunedisorders, such as systemic lupus erythematosus and sclerodermatitis,chemical contact or allergies. Another autoimmune disorder associatedwith fibroproliferative characteristics is rheumatoid arthritis.

Eye diseases associated with a fibroproliferative condition includeretinal reattachment surgery accompanying proliferativevitreoretinopathy, cataract extraction with intraocular lensimplantation, and post-glaucoma drainage surgery and are associated withTGF-β1 overproduction.

Fibrotic diseases associated with TGF-β1 overproduction can be dividedinto chronic conditions, such as fibrosis of the kidney, lung and liver,and more acute conditions, such as dermal scarring and restenosis(Chamberlain, J. Cardiovascular Drug Reviews, 19(4): 329-344). Synthesisand secretion of TGF-β1 by tumour cells can also lead to immunesuppression, as seen in patients with aggressive brain or breast tumours(Arteaga, et al. (1993) J. Clin. Invest. 92: 2569-2576). The course ofleishmanial infection in mice is drastically altered by TGF-β1(Barral-Netto, et al. (1992) Science 257: 545-547). TGF-β1 exacerbatedthe disease, whereas TGF-β1 antibodies halted the progression of thedisease in genetically susceptible mice. Genetically resistant micebecame susceptible to leishmanial infection upon administration ofTGF-β1.

The profound effects on extracellular matrix deposition have beenreviewed (Rocco and Ziyadeh (1991) in Contemporary Issues in Nephrologyv.23, Hormones, autocoids and the kidney. ed. Jay Stein, ChurchillLivingston, New York pp. 391-410; Roberts, et al. (1988) Rec. Prog.Hormone Res. 44: 157-197) and include stimulation of the synthesis andinhibition of the degradation of extracellular matrix components. Sincethe structural and filtration properties of the glomerulus are largelydetermined by the extracellular matrix composition of the mesangium andglomerular membrane, it is not surprising that TGF-β1 has profoundeffects on the kidney. The accumulation of mesangial matrix inproliferative glomerulonephritis (Border, et al., (1990) Kidney Int. 37:689-695) and diabetic nephropathy (Mauer, et al. (1984) J. Clin. Invest.74: 1143-1155) are clear and dominant pathological features of thediseases. TGF-β1 levels are elevated in human diabeticglomerulosclerosis (advanced neuropathy) (Yamamoto, et al. (1993) Proc.Natl. Acad. Sci. 90: 1814-1818). TGF-β1 is an important mediator in thegenesis of renal fibrosis in a number of animal models (Phan, et al.(1990) Kidney Int. 37: 426; Okuda, et al. (1990) J. Clin. Invest. 86:453). Suppression of experimentally induced glomerulonephritis in ratshas been demonstrated by antiserum against TGF-β1 (Border, et al. (1990)Nature 346: 371) and by an extracellular matrix protein, decorin, whichcan bind TGF-β1 (Border, et al. (1992) Nature 360: 361-363).

Excessive TGF-β1 leads to dermal scar-tissue formation. NeutralisingTGF-β1 antibodies injected into the margins of healing wounds in ratshas been shown to inhibit scarring without interfering with the rate ofwound healing or the tensile strength of the wound (Shah, et al. (1992)Lancet 339: 213-214). At the same time there was reduced angiogenesis, areduced number of macrophages and monocytes in the wound, and a reducedamount of disorganised collagen fibre deposition in the scar tissue.

TGF-β1 may be a factor in the progressive thickening of the arterialwall which results from the proliferation of smooth muscle cells anddeposition of extracellular matrix in the artery after balloonangioplasty. The diameter of the restenosed artery may be reduced by 90%by this thickening, and since most of the reduction in diameter is dueto extracellular matrix rather than smooth muscle cell bodies, it may bepossible to reopen these vessels to 50% simply by reducing excessiveextracellular matrix deposition. In undamaged pig arteries transfectedin vivo with a TGF-β1 gene, TGF-β1 gene expression was associated withboth extracellular matrix synthesis and hyperplasia (Nebel, et al.(1993) Proc. Natl. Acad. Sci USA 90: 10759-10763). The TGF-β1-inducedhyperplasia was not as extensive as that induced with PDGF-BB, but theextracellular matrix was more extensive with TGF-β1 transfectants. Noextracellular matrix deposition was associated with hyperplasia inducedby FGF-1 (a secreted form of FGF) in this gene transfer pig model (Nabel(1993) Nature 362: 844-846).

There are various types of cancer where TGF-β1 produced by the tumourmay be deleterious. MATLyLu rat prostate cancer cells (Steiner andBarrack (1992) Mol. Endocrinol 6: 15-25) and MCF-7 human breast cancercells (Arteaga, et al. (1993) Cell Growth and Differ. 4: 193-201) becamemore tumorigenic and metastatic after transfection with a vectorexpressing the mouse TGF-β1. TGF-β1 has been associated withangiogenesis, metastasis and poor prognosis in human prostate andadvanced intestinal cancer (Wikstrom, P., et al. (1988) Prostate 37;19-29; Saito, H., et al. (1999) Cancer 86: 1455-1462). In breast cancer,a poor prognosis is associated with elevated TGF-β (Dickson, et al.(1987) Proc. Natl. Acad. Sci. USA 84: 837-841; Kasid, et al. (1987)Cancer Res. 47: 5733-5738; Daly, et al. (1990) J. Cell Biochem. 43:199-211; Barrett-Lee, et al. (1990) Br. J. Cancer 61: 612-617; King, etal (1989) J. Steroid Biochem. 34: 133-138; Welch, et al (1990) Proc.Natl. Acad. Sci USA 87: 7678-7682; Walker et al. (1992) Eur. J. Cancer238: 641-644), and induction of TGF-β1 by tamoxifen treatment (Butta, etal. (1992) Cancer Res. 52: 4261-4264) has been associated with failureof tamoxifen treatment for breast cancer (Thompson, et al. (1991) Br. J.Cancer 63: 609-614). Anti-TGF-β1 antibodies inhibit the growth ofMDA-231 human breast cancer cells in athymic mice (Arteaga, et al.(1993) J. Clin. Invest. 92: 2569-2576), a treatment which is correlatedwith an increase in natural killer cell activity in the spleen. CHOcells transfected with latent TGF-β1 also showed decreased NK activityand increased tumour growth in nude mice (Wallick, et al. (1990) J. Exp.Med. 172: 177-1784). Thus, TGF-β secreted by breast tumours may causeendocrine immune suppression. High plasma concentrations of TGF-β1 showa poor prognosis for advanced breast cancer patients (Anscher, et al.(1993) N. Engl. J. Med. 328: 1592-1598). Patients with high circulatingTGF-β before high dose chemotherapy and autologous bone marrowtransplantation are at high risk of a hepatic veno-occlusive disease(15-50% of all patients with a mortality rate up to 50%) and idiopathicinterstitial pneumonitis (40 to 60% of all patients). The implication ofthese findings is 1) that elevated plasma levels of TGF-β1 can be usedto identify at-risk patients and 2) that reduction of TGF-β1 candecrease the morbidity and mortality of these common treatments forbreast cancer patients.

Many malignant cells secrete transforming growth factor β (TGF-β), apotent immunosuppressant, suggesting that TGF-β production may representa significant tumour escape mechanism from host immunosurveillance.Establishment of a leukocyte sub-population with a disrupted TGF-βsignalling pathway in the tumour-bearing host offers a powerful measurefor immunotherapy of cancer. A transgenic animal model with a disruptedTGF-β signalling pathway in T cells is capable of eradicating a normallylethal TGF-β-overexpressing lymphoma tumour, EL4 (Gorelik and Flavell,(2001) Nature Medicine 7 (10): 1118-1122). Downregulation of TGF-βsecretion in tumour cells results in restoration of immunogenicity inthe host, while T-cell insensitivity to TGF-β results in accelerateddifferentiation and autoimmunity, elements of which may be required inorder to combat self-antigen-expressing tumours in a tolerised host. Theimmunosuppressive effects of TGF-β have also been implicated in asub-population of HIV patients with lower than predicted immune responsebased on their CD4/CD8 T cell counts (Garba, et al., J. Immunology(2002) 168: 2247-2254). A TGF-β-neutralising antibody was capable ofreversing the effect in culture, indicating that TGF-β signallingpathway inhibitors may be suitable in reversing the immune suppressionpresent in this subset of HIV patients.

During the earliest stages of carcinogenesis, TGF-β1 can act as a potenttumour suppressor and may mediate the actions of some chemopreventiveagents. At a certain point during the development and progression ofmalignant neoplasms, tumour cells appear to escape from TGF-β-dependentgrowth inhibition in parallel with the appearance of biologically activeTGF-β in the microenvironment. The dual tumour suppression/tumourpromotion roles of TGF-β have been most clearly elucidated in atransgenic system overexpressing TGF-β in keratinocytes. While thetransgenics were more resistant to formation of benign skin lesions, therate of metastatic conversion in the transgenics was dramaticallyincreased (Cui, et al, (1996) Cell 86(4): 531-42). The production ofTGF-β1 by malignant cells in primary tumours appears to increase withadvancing stages of tumour progression. Studies in many of the majorepithelial cancers suggest that the increased production of TGF-β byhuman cancers occurs as a relatively late event during tumourprogression. Furthermore, this tumour-associated TGF-β provides thetumour cells with a selective advantage and promotes tumour progression.The effects of TGF-β on cell-cell and cell-stroma interactions resultsin a greater propensity for invasion and metastasis. Tumour-associatedTGF-β may allow tumour cells to escape from immunosurveillance since itis a potent inhibitor of the clonal expansion of activated lymphocytes.TGF-β has also been shown to inhibit the production of angiostatin.Cancer therapeutic modalities, such as radiation therapy andchemotherapy, induce the production of activated TGF-β in the tumour,thereby selecting outgrowth of malignant cells that are resistant toTGF-β growth inhibitory effects. Thus, these anticancer treatmentsincrease the risk and hasten the development of tumours with enhancedgrowth and invasiveness. In this situation, agents targetingTGF-β-mediated signal transduction might be a very effective therapeuticstrategy. The resistance of tumour cells to TGF-β has been shown tonegate many of the cytotoxic effects of radiation therapy andchemotherapy, and the treatment-dependent activation of TGF-β in thestroma may even be detrimental as it makes the microenvironment moreconducive to tumour progression and contributes to tissue damage leadingto fibrosis. The development of TGF-β signal transduction inhibitors islikely to benefit the treatment of advanced cancer alone and incombination with other therapies.

The compounds are suitable for the treatment of cancer and otherconditions influenced by TGF-β by inhibiting TGF-β in a patient in needthereof by administration of the compound(s) to the patient. TGF-β isalso suitable against atherosclerotic (T. A. McCaffrey: TGF-βs and TGF-βReceptors in Atherosclerosis: Cytokine and Growth Factor Reviews 2000,11, 103-114) and Alzheimer's diseases (Masliah, E.; Ho, G.; Wyss-Coray,T.: Functional Role of TGF-β in Alzheimer's Disease MicrovascularInjury: Lessons from Transgenic Mice: Neurochemistry International 2001,39, 393-400).

It has been found that the compounds according to the invention andsalts thereof have very valuable pharmacological properties while beingwell tolerated.

In particular, they exhibit TGF receptor I kinase-inhibiting properties.

The compounds according to the invention preferably exhibit anadvantageous biological activity, which can easily be demonstrated inenzyme-based assays, for example assays as described herein. In suchenzyme-based assays, the compounds according to the invention preferablyexhibit and cause an inhibiting effect, which is usually documented byIC₅₀ values in a suitable range, preferably in the micromolar range andmore preferably in the nanomolar range.

As discussed herein, these signalling pathways are relevant for variousdiseases. Accordingly, the compounds according to the invention areuseful in the prophylaxis and/or treatment of diseases that aredependent on the said signalling pathways by interaction with one ormore of the said signalling pathways.

The present invention therefore relates to compounds according to theinvention as promoters or inhibitors, preferably as inhibitors, of thesignalling pathways described herein. The invention therefore preferablyrelates to compounds according to the invention as promoters orinhibitors, preferably as inhibitors, of the TGF signalling pathway.

The present invention furthermore relates to the use of one or morecompounds according to the invention in the treatment and/or prophylaxisof diseases, preferably the diseases described herein, that are caused,mediated and/or propagated by an increased TGF activity.

The present invention therefore relates to compounds according to theinvention as medicaments and/or medicament active ingredients in thetreatment and/or prophylaxis of the said diseases and to the use ofcompounds according to the invention for the preparation of apharmaceutical for the treatment and/or prophylaxis of the said diseasesas well as to a method for the treatment of the said diseases comprisingthe administration of one or more compounds according to the inventionto a patient in need of such an administration.

The host or patient can belong to any mammalian species, for example aprimate species, particularly humans; rodents, including mice, rats andhamsters; rabbits; horses, cows, dogs, cats, etc. Animal models are ofinterest for experimental investigations, providing a model fortreatment of a human disease.

The susceptibility of a particular cell to treatment with the compoundsaccording to the invention can be determined by in-vitro testing.Typically, a culture of the cell is combined with a compound accordingto the invention at various concentrations for a period of time which issufficient to allow the active agents to induce cell death or to inhibitmigration, usually between about one hour and one week. In-vitro testingcan be carried out using cultivated cells from a biopsy sample. Theviable cells remaining after the treatment are then counted.

The dose varies depending on the specific compound used, the specificdisease, the patient status, etc. A therapeutic dose is typicallysufficient considerably to reduce the undesired cell population in thetarget tissue while the viability of the patient is maintained. Thetreatment is generally continued until a considerable reduction hasoccurred, for example an at least about 50% reduction in the cellburden, and may be continued until essentially no more undesired cellsare detected in the body.

For identification of a signal transduction pathway and for detection ofinteractions between various signal transduction pathways, variousscientists have developed suitable models or model systems, for examplecell culture models (for example Khwaja et al., EMBO, 1997, 16, 2783-93)and models of transgenic animals (for example White et al., Oncogene,2001, 20, 7064-7072). For the determination of certain stages in thesignal transduction cascade, interacting compounds can be utilised inorder to modulate the signal (for example Stephens et al., BiochemicalJ., 2000, 351, 95-105). The compounds according to the invention canalso be used as reagents for testing kinase-dependent signaltransduction pathways in animals and/or cell culture models or in theclinical diseases mentioned in this application.

Measurement of the kinase activity is a technique which is well known tothe person skilled in the art. Generic test systems for thedetermination of the kinase activity using substrates, for examplehistone (for example Alessi et al., FEBS Lett. 1996, 399, 3, pages333-338) or the basic myelin protein, are described in the literature(for example Campos-González, R. and Glenney, Jr., J. R. 1992, J. Biol.Chem. 267, page 14535).

For the identification of kinase inhibitors, various assay systems areavailable. In the scintillation proximity assay (Sorg et al., J. of.Biomolecular Screening, 2002, 7, 11-19) and the flashplate assay, theradioactive phosphorylation of a protein or peptide as substrate withγATP is measured. In the presence of an inhibitory compound, a decreasedradioactive signal, or none at all, is detectable. Furthermore,homogeneous time-resolved fluorescence resonance energy transfer(HTR-FRET) and fluorescence polarisation (FP) technologies are suitableas assay methods (Sills et al., J. of Biomolecular Screening, 2002,191-214).

Other non-radioactive ELISA assay methods use specificphospho-antibodies (phospho-ABs). The phospho-AB binds only thephosphorylated substrate. This binding can be detected bychemiluminescence using a second peroxidase-conjugated anti-sheepantibody (Ross et al., 2002, Biochem. J., just about to be published,manuscript BJ20020786).

PRIOR ART

Triazolo-1,5-benzodiazepines are known from DE 2 318 673.

L. Kosychova et al. in Chemistry of Heterocyclic Compounds, Vol. 40,811-815 (2004) describe other5,6-dihydro-4H-1,2,4-triazolo-a]-1,5-benzodiazepines for combatingtumours.

V. Ambrogi et al. in J. Heterocyclic Chem. 31, 1349-1352 (1994) describesulfur-containing 4,5-dihydro-s-triazolo[3,4d]-1,5-benzothiazepinederivatives.

V. Ambrogi et al. in II Farmaco 48, 665-676 (1993) describe1,4-benzothiazines and 1,5-benzothiazepines having an action on thecentral nervous system.

Other triazole derivatives are disclosed as TGF-beta inhibitors in WO03/042211 A1.

Still other triazole derivatives are known as TGF-beta inhibitors fromWO 2004/026307 A1.

Bicyclic pyrrole derivatives are described as TGF-beta inhibitors in WO02/094833.

SUMMARY OF THE INVENTION

The invention relates to compounds of the formula I

in which

-   -   R¹ denotes H, A, OH, OA, NO₂, NH₂, NHA, NA₂, Hal, CN, A-COO,        COOH, COOA or CONR⁴R⁵,    -   R², R³ each, independently of one another, denote H, A, alkenyl        having 2-6 C atoms, alkynyl having 2-6 C atoms or Hal,    -   R⁴, R⁵ each, independently of one another, denote H or A,    -   A denotes unbranched or branched alkyl having 1, 2, 3, 4, 5, 6,        7, 8, 9 or 10 C atoms, in which 1-7 H atoms may be replaced by        F,    -   Hal denotes F, Cl, Br or I,        and pharmaceutically usable derivatives, solvates, salts,        tautomers and stereoisomers thereof, including mixtures thereof        in all ratios.

The invention also relates to the optically active forms(stereoisomers), the enantiomers, the racemates, the diastereomers andthe hydrates and solvates of these compounds. The term solvates of thecompounds is taken to mean adductions of inert solvent molecules ontothe compounds which form owing to their mutual attractive force.Solvates are, for example, mono- or dihydrates or alkoxides.

The term pharmaceutically usable derivatives is taken to mean, forexample, the salts of the compounds according to the invention and alsoso-called prodrug compounds.

The term prodrug derivatives is taken to mean compounds according to theinvention which have been modified by means of, for example, alkyl oracyl groups, sugars or oligopeptides and which are rapidly cleaved inthe organism to form the effective compounds according to the invention.These also include biodegradable polymer derivatives of the compoundsaccording to the invention, as described, for example, in Int. J. Pharm.115, 61-67 (1995).

The expression “effective amount” denotes the amount of a medicament orof a pharmaceutical active ingredient which causes in a tissue, system,animal or human a biological or medical response which is sought ordesired, for example, by a researcher or physician.

In addition, the expression “therapeutically effective amount” denotesan amount which, compared with a corresponding subject who has notreceived this amount, has the following consequence:

improved treatment, healing, prevention or elimination of a disease,syndrome, condition, complaint, disorder or side-effects or also thereduction in the advance of a disease, complaint or disorder.

The expression “therapeutically effective amount” also encompasses theamounts which are effective for increasing normal physiologicalfunction.

The invention also relates to the use of mixtures of the compoundsaccording to the invention, for example mixtures of two diastereomers,for example in the ratio 1:1, 1:2, 1:3, 1:4, 1:5, 1:10, 1:100 or 1:1000.

These are particularly preferably mixtures of stereoisomeric compounds.

The invention relates to the compounds of the formula I and saltsthereof and to a process for the preparation of compounds of the formulaI and pharmaceutically usable derivatives, salts, solvates, tautomersand stereoisomers thereof, characterised in that

a) a compound of the formula II

in which R¹ and R² has the meaning indicated in Claim 1,

is reacted with a compound of the formula III

in which R³ has the meaning indicated in Claim 1,

or

b) a radical R¹ is converted into another radical R¹ by cleaving anether,

and/or

a base or acid of the formula I is converted into one of its salts.

Above and below, the radicals R¹, R² and R³ have the meanings indicatedfor the formula I, unless expressly indicated otherwise.

A denotes alkyl, is unbranched (linear) or branched, and has 1, 2, 3, 4,5, 6, 7, 8, 9 or 10 C atoms. A preferably denotes methyl, furthermoreethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl,furthermore also pentyl, 1-, 2- or 3-methylbutyl, 1,1- , 1,2- or2,2-dimethylpropyl, 1-ethylpropyl, hexyl, 1-, 2-, 3- or 4-methylpentyl,1,1-, 1,2-, 1,3-, 2,2-, 2,3- or 3,3-dimethylbutyl, 1- or 2-ethylbutyl,1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, 1,1,2- or1,2,2-trimethylpropyl, further preferably, for example, trifluoromethyl.

A very particularly preferably denotes alkyl having 1, 2, 3, 4, 5 or 6 Catoms, preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl,sec-butyl, tert-butyl, pentyl, hexyl, trifluoromethyl, pentafluoroethylor 1,1,1-trifluoroethyl.

R¹ preferably denotes H, OH, OA, Hal, CN or A-COO.

R² preferably denotes H or A.

R³ preferably denotes A.

Throughout the invention, all radicals which occur more than once may beidentical or different, i.e. are independent of one another.

The compounds of the formula I may have one or more chiral centres andtherefore occur in various stereoisomeric forms. The formula Iencompasses all these forms.

Accordingly, the invention relates, in particular, to the compounds ofthe formula I in which at least one of the said radicals has one of thepreferred meanings indicated above. Some preferred groups of compoundscan be expressed by the following sub-formulae Ia to Id, which conformto the formula I and in which the radicals not designated in greaterdetail have the meaning indicated for the formula I, but in which

-   -   in Ia R¹ denotes H, OH, OA, Hal, CN or A-COO;    -   in Ib R² denotes H or A;    -   in Ic R³ denotes A;    -   in Id R¹ denotes H, OH, OA, Hal, CN or A-COO,        -   R² denotes H or A,        -   R³ R denotes A,        -   A denotes unbranched or branched alkyl having 1, 2, 3, 4, 5            or 6 C atoms, in which 1-5 H atoms may be replaced by F,        -   Hal denotes F, Cl, Br or I;            and pharmaceutically usable derivatives, salts, solvates,            tautomers and stereoisomers thereof, including mixtures            thereof in all ratios.

The compounds of the formula I and also the starting materials for theirpreparation are, in addition, prepared by methods known per se, asdescribed in the literature (for example in the standard works, such asHouben-Weyl, Methoden der organischen Chemie [Methods of OrganicChemistry], Georg-Thieme-Verlag, Stuttgart), to be precise underreaction conditions which are known and suitable for the said reactions.Use can also be made here of variants known per se which are notmentioned here in greater detail.

The starting compounds of the formulae II and III are generally known.If they are novel, however, they can be prepared by methods known perse.

Compounds of the formula I can preferably be obtained by reacting acompound of the formula II with a compound of the formula III. Thereaction is carried out in an inert solvent. Depending on the conditionsused, the reaction time is between a few minutes and 14 days, thereaction temperature is between about 0° and 160°, normally between 20°and 150°, in particular between about 80° and about 130°.

Suitable inert solvents are, for example, hydrocarbons, such as hexane,petroleum ether, benzene, toluene or xylene; chlorinated hydrocarbons,such as trichloroethylene, 1,2-dichloroethane, carbon tetrachloride,chloroform or dichloromethane; alcohols, such as methanol, ethanol,isopropanol, n-propanol, n-butanol or tert-butanol; ethers, such asdiethyl ether, diisopropyl ether, tetrahydrofuran (THF) or dioxane;glycol ethers, such as ethylene glycol monomethyl or monoethyl ether,ethylene glycol dimethyl ether (diglyme); ketones, such as acetone orbutanone; amides, such as acetamide, dimethylacetamide,1-methylpyrrolidinone (NMP) or dimethylformamide (DMF); nitriles, suchas acetonitrile; sulfoxides, such as dimethyl sulfoxide (DMSO); carbondisulfide; carboxylic acids, such as formic acid or acetic acid; nitrocompounds, such as nitromethane or nitrobenzene; esters, such as ethylacetate, or mixtures of the said solvents.

Particular preference is given to n-butanol, NMP and/or DMF.

Suitable for the cleavage of ethers is treatment with boron tribromideunder standard conditions.

Pharmaceutical Salts and Other Forms

The said compounds according to the invention can be used in their finalnon-salt form. On the other hand, the present invention also encompassesthe use of these compounds in the form of their pharmaceuticallyacceptable salts, which can be derived from various organic andinorganic acids and bases by procedures known in the art.Pharmaceutically acceptable salt forms of the compounds according to theinvention are for the most part prepared by conventional methods. If thecompound according to the invention contains a carboxyl group, one ofits suitable salts can be formed by reacting the compound with asuitable base to give the corresponding base-addition salt. Such basesare, for example, alkali metal hydroxides, including potassiumhydroxide, sodium hydroxide and lithium hydroxide; alkaline earth metalhydroxides, such as barium hydroxide and calcium hydroxide; alkali metalalkoxides, for example potassium ethoxide and sodium propoxide; andvarious organic bases, such as piperidine, diethanolamine andN-methylglutamine. The aluminium salts of the compounds according to theinvention are likewise included. In the case of certain compoundsaccording to the invention, acid-addition salts can be formed bytreating these compounds with pharmaceutically acceptable organic andinorganic acids, for example hydrogen halides, such as hydrogenchloride, hydrogen bromide or hydrogen iodide, other mineral acids andcorresponding salts thereof, such as sulfate, nitrate or phosphate andthe like, and alkyl- and monoarylsulfonates, such as ethanesulfonate,toluenesulfonate and benzenesulfonate, and other organic acids andcorresponding salts thereof, such as acetate, trifluoroacetate,tartrate, maleate, succinate, citrate, benzoate, salicylate, ascorbateand the like. Accordingly, pharmaceutically acceptable acid-additionsalts of the compounds according to the invention include the following:acetate, adipate, alginate, arginate, aspartate, benzoate,benzenesulfonate (besylate), bisulfate, bisulfite, bromide, butyrate,camphorate, camphorsulfonate, caprylate, chloride, chlorobenzoate,citrate, cyclopentanepropionate, digluconate, dihydrogenphosphate,dinitrobenzoate, dodecylsulfate, ethanesulfonate, fumarate, galacterate(from mucic acid), galacturonate, glucoheptanoate, gluconate, glutamate,glycerophosphate, hemisuccinate, hemisulfate, heptanoate, hexanoate,hippurate, hydrochloride, hydrobromide, hydroiodide,2-hydroxyethanesulfonate, iodide, isethionate, isobutyrate, lactate,lactobionate, malate, maleate, malonate, mandelate, metaphosphate,methanesulfonate, methylbenzoate, monohydrogenphosphate,2-naphthalenesulfonate, nicotinate, nitrate, oxalate, oleate, palmoate,pectinate, persulfate, phenylacetate, 3-phenylpropionate, phosphate,phosphonate, phthalate, but this does not represent a restriction.

Furthermore, the base salts of the compounds according to the inventioninclude aluminium, ammonium, calcium, copper, iron(III), iron(II),lithium, magnesium, manganese(III), manganese(II), potassium, sodium andzinc salts, but this is not intended to represent a restriction. Of theabove-mentioned salts, preference is given to ammonium; the alkali metalsalts sodium and potassium, and the alkaline earth metal salts calciumand magnesium. Salts of the compounds according to the invention whichare derived from pharmaceutically acceptable organic non-toxic basesinclude salts of primary, secondary and tertiary amines, substitutedamines, also including naturally occurring substituted amines, cyclicamines, and basic ion exchanger resins, for example arginine, betaine,caffeine, chloroprocaine, choline, N,N′-dibenzylethylenediamine(benzathine), dicyclohexylamine, diethanolamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lidocaine, lysine,meglumine, N-methyl-D-glucamine, morpholine, piperazine, piperidine,polyamine resins, procaine, purines, theobromine, triethanolamine,triethylamine, trimethylamine, tripropylamine andtris(hydroxymethyl)methylamine (tromethamine), but this is not intendedto represent a restriction.

Compounds of the present invention which contain basicnitrogen-containing groups can be quaternised using agents such as(C₁-C₄)alkyl halides, for example methyl, ethyl, isopropyl andtert-butyl chloride, bromide and iodide; di(C₁-C₄)alkyl sulfates, forexample dimethyl, diethyl and diamyl sulfate; (C₁₀-C₁₈)alkyl halides,for example decyl, dodecyl, lauryl, myristyl and stearyl chloride,bromide and iodide; and aryl(C₁-C₄)alkyl halides, for example benzylchloride and phenethyl bromide. Both water- and oil-soluble compoundsaccording to the invention can be prepared using such salts.

The above-mentioned pharmaceutical salts which are preferred includeacetate, trifluoroacetate, besylate, citrate, fumarate, gluconate,hemisuccinate, hippurate, hydrochloride, hydrobromide, isethionate,mandelate, meglumine, nitrate, oleate, phosphonate, pivalate, sodiumphosphate, stearate, sulfate, sulfosalicylate, tartrate, thiomalate,tosylate and tromethamine, but this is not intended to represent arestriction.

The acid-addition salts of basic compounds of the compounds according tothe invention are prepared by bringing the free base form into contactwith a sufficient amount of the desired acid, causing the formation ofthe salt in a conventional manner. The free base can be regenerated bybringing the salt form into contact with a base and isolating the freebase in a conventional manner. The free base forms differ in a certainrespect from the corresponding salt forms thereof with respect tocertain physical properties, such as solubility in polar solvents; forthe purposes of the invention, however, the salts otherwise correspondto the respective free base forms thereof.

As mentioned, the pharmaceutically acceptable base-addition salts of thecompounds according to the invention are formed with metals or amines,such as alkali metals and alkaline earth metals or organic amines.Preferred metals are sodium, potassium, magnesium and calcium. Preferredorganic amines are N,N′-dibenzylethylenediamine, chloroprocaine,choline, diethanolamine, ethylenediamine, N-methyl-D-glucamine andprocaine.

The base-addition salts of acidic compounds according to the inventionare prepared by bringing the free acid form into contact with asufficient amount of the desired base, causing the formation of the saltin a conventional manner. The free acid can be regenerated by bringingthe salt form into contact with an acid and isolating the free acid in aconventional manner. The free acid forms differ in a certain respectfrom the corresponding salt forms thereof with respect to certainphysical properties, such as solubility in polar solvents; for thepurposes of the invention, however, the salts otherwise correspond tothe respective free acid forms thereof.

If a compound according to the invention contains more than one groupwhich is capable of forming pharmaceutically acceptable salts of thistype, the invention also encompasses multiple salts. Typical multiplesalt forms include, for example, bitartrate, diacetate, difumarate,dimeglumine, diphosphate, disodium and trihydrochloride, but this is notintended to represent a restriction.

With regard to that stated above, it can be seen that the expression“pharmaceutically acceptable salt” in the present connection is taken tomean an active ingredient which comprises a compound according to theinvention in the form of one of its salts, in particular if this saltform imparts improved pharmacokinetic properties on the activeingredient compared with the free form of the active ingredient or anyother salt form of the active ingredient used earlier. Thepharmaceutically acceptable salt form of the active ingredient can alsoprovide this active ingredient for the first time with a desiredpharmacokinetic property which it did not have earlier and can even havea positive influence on the pharmacodynamics of this active ingredientwith respect to its therapeutic efficacy in the body.

The invention furthermore relates to medicaments comprising at least onecompound according to the invention and/or pharmaceutically usablederivatives, solvates and stereoisomers thereof, including mixturesthereof in all ratios, and optionally excipients and/or adjuvants.

Pharmaceutical formulations can be administered in the form of dosageunits which comprise a predetermined amount of active ingredient perdosage unit. Such a unit can comprise, for example, 0.5 mg to 1 g,preferably 1 mg to 700 mg, particularly preferably 5 mg to 100 mg, of acompound according to the invention, depending on the condition treated,the method of administration and the age, weight and condition of thepatient, or pharmaceutical formulations can be administered in the formof dosage units which comprise a predetermined amount of activeingredient per dosage unit. Preferred dosage unit formulations are thosewhich comprise a daily dose or part-dose, as indicated above, or acorresponding fraction thereof of an active ingredient. Furthermore,pharmaceutical formulations of this type can be prepared using a processwhich is generally known in the pharmaceutical art.

Pharmaceutical formulations can be adapted for administration via anydesired suitable method, for example by oral (including buccal orsublingual), rectal, nasal, topical (including buccal, sublingual ortransdermal), vaginal or parenteral (including subcutaneous,intramuscular, intravenous or intradermal) methods. Such formulationscan be prepared using all processes known in the pharmaceutical art by,for example, combining the active ingredient with the excipient(s) oradjuvant(s).

Pharmaceutical formulations adapted for oral administration can beadministered as separate units, such as, for example, capsules ortablets; powders or granules; solutions or suspensions in aqueous ornon-aqueous liquids; edible foams or foam foods; or oil-in-water liquidemulsions or water-in-oil liquid emulsions.

Thus, for example, in the case of oral administration in the form of atablet or capsule, the active-ingredient component can be combined withan oral, non-toxic and pharmaceutically acceptable inert excipient, suchas, for example, ethanol, glycerol, water and the like. Powders areprepared by comminuting the compound to a suitable fine size and mixingit with a pharmaceutical excipient comminuted in a similar manner, suchas, for example, an edible carbohydrate, such as, for example, starch ormannitol. A flavour, preservative, dispersant and dye may likewise bepresent.

Capsules are produced by preparing a powder mixture as described aboveand filling shaped gelatine shells therewith. Glidants and lubricants,such as, for example, highly disperse silicic acid, talc, magnesiumstearate, calcium stearate or polyethylene glycol in solid form, can beadded to the powder mixture before the filling operation. A disintegrantor solubiliser, such as, for example, agar-agar, calcium carbonate orsodium carbonate, may likewise be added in order to improve theavailability of the medicament after the capsule has been taken.

In addition, if desired or necessary, suitable binders, lubricants anddisintegrants as well as dyes can likewise be incorporated into themixture. Suitable binders include starch, gelatine, natural sugars, suchas, for example, glucose or beta-lactose, sweeteners made from maize,natural and synthetic rubber, such as, for example, acacia, tragacanthor sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes,and the like. The lubricants used in these dosage forms include sodiumoleate, sodium stearate, magnesium stearate, sodium benzoate, sodiumacetate, sodium chloride and the like. The disintegrants include,without being restricted thereto, starch, methylcellulose, agar,bentonite, xanthan gum and the like. The tablets are formulated by, forexample, preparing a powder mixture, granulating or dry-pressing themixture, adding a lubricant and a disintegrant and pressing the entiremixture to give tablets. A powder mixture is prepared by mixing thecompound comminuted in a suitable manner with a diluent or a base, asdescribed above, and optionally with a binder, such as, for example,carboxymethylcellulose, an alginate, gelatine or polyvinylpyrrolidone, adissolution retardant, such as, for example, paraffin, an absorptionaccelerator, such as, for example, a quaternary salt, and/or anabsorbant, such as, for example, bentonite, kaolin or dicalciumphosphate. The powder mixture can be granulated by wetting it with abinder, such as, for example, syrup, starch paste, acadia mucilage orsolutions of cellulose or polymer materials and pressing it through asieve. As an alternative to granulation, the powder mixture can be runthrough a tableting machine, giving lumps of non-uniform shape, whichare broken up to form granules. The granules can be lubricated byaddition of stearic acid, a stearate salt, talc or mineral oil in orderto prevent sticking to the tablet casting moulds. The lubricated mixtureis then pressed to give tablets. The compounds according to theinvention can also be combined with a free-flowing inert excipient andthen pressed directly to give tablets without carrying out thegranulation or dry-pressing steps. A transparent or opaque protectivelayer consisting of a shellac sealing layer, a layer of sugar or polymermaterial and a gloss layer of wax may be present. Dyes can be added tothese coatings in order to be able to differentiate between differentdosage units.

Oral liquids, such as, for example, solution, syrups and elixirs, can beprepared in the form of dosage units so that a given quantity comprisesa prespecified amount of the compound. Syrups can be prepared bydissolving the compound in an aqueous solution with a suitable flavour,while elixirs are prepared using a non-toxic alcoholic vehicle.Suspensions can be formulated by dispersion of the compound in anon-toxic vehicle. Solubilisers and emulsifiers, such as, for example,ethoxylated isostearyl alcohols and polyoxyethylene sorbitol ethers,preservatives, flavour additives, such as, for example, peppermint oilor natural sweeteners or saccharin, or other artificial sweeteners andthe like, can likewise be added.

The dosage unit formulations for oral administration can, if desired, beencapsulated in microcapsules. The formulation can also be prepared insuch a way that the release is extended or retarded, such as, forexample, by coating or embedding of particulate material in polymers,wax and the like.

The compounds according to the invention and salts, solvates andphysiologically functional derivatives thereof can also be administeredin the form of liposome delivery systems, such as, for example, smallunilamellar vesicles, large unilamellar vesicles and multilamellarvesicles. Liposomes can be formed from various phospholipids, such as,for example, cholesterol, stearylamine or phosphatidylcholines.

The compounds according to the invention and the salts, solvates andphysiologically functional derivatives thereof can also be deliveredusing monoclonal antibodies as individual carriers to which the compoundmolecules are coupled. The compounds can also be coupled to solublepolymers as targeted medicament carriers. Such polymers may encompasspolyvinylpyrrolidone, pyran copolymer,polyhydroxypropylmethacrylamidophenol, polyhydroxyethylaspartamidophenolor polyethylene oxide polylysine, substituted by palmitoyl radicals. Thecompounds may furthermore be coupled to a class of biodegradablepolymers which are suitable for achieving controlled release of amedicament, for example polylactic acid, poly-epsilon-caprolactone,polyhydroxybutyric acid, polyorthoesters, polyacetals,polydihydroxypyrans, polycyanoacrylates and crosslinked or amphipathicblock copolymers of hydrogels.

Pharmaceutical formulations adapted for transdermal administration canbe administered as independent plasters for extended, close contact withthe epidermis of the recipient. Thus, for example, the active ingredientcan be delivered from the plaster by iontophoresis, as described ingeneral terms in Pharmaceutical Research, 3(6), 318 (1986).

Pharmaceutical compounds adapted for topical administration can beformulated as ointments, creams, suspensions, lotions, powders,solutions, pastes, gels, sprays, aerosols or oils.

For the treatment of the eye or other external tissue, for example mouthand skin, the formulations are preferably applied as topical ointment orcream. In the case of formulation to give an ointment, the activeingredient can be employed either with a paraffinic or a water-misciblecream base. Alternatively, the active ingredient can be formulated togive a cream with an oil-in-water cream base or a water-in-oil base.

Pharmaceutical formulations adapted for topical application to the eyeinclude eye drops, in which the active ingredient is dissolved orsuspended in a suitable carrier, in particular an aqueous solvent.

Pharmaceutical formulations adapted for topical application in the mouthencompass lozenges, pastilles and mouthwashes.

Pharmaceutical formulations adapted for rectal administration can beadministered in the form of suppositories or enemas.

Pharmaceutical formulations adapted for nasal administration in whichthe carrier substance is a solid comprise a coarse powder having aparticle size, for example, in the range 20-500 microns, which isadministered in the manner in which snuff is taken, i.e. by rapidinhalation via the nasal passages from a container containing the powderheld close to the nose.

Suitable formulations for administration as nasal spray or nose dropswith a liquid as carrier substance encompass active-ingredient solutionsin water or oil.

Pharmaceutical formulations adapted for administration by inhalationencompass finely particulate dusts or mists, which can be generated byvarious types of pressurised dispensers with aerosols, nebulisers orinsufflators.

Pharmaceutical formulations adapted for vaginal administration can beadministered as pessaries, tampons, creams, gels, pastes, foams or sprayformulations.

Pharmaceutical formulations adapted for parenteral administrationinclude aqueous and non-aqueous sterile injection solutions comprisingantioxidants, buffers, bacteriostatics and solutes, by means of whichthe formulation is rendered isotonic with the blood of the recipient tobe treated; and aqueous and non-aqueous sterile suspensions, which maycomprise suspension media and thickeners. The formulations can beadministered in single-dose or multidose containers, for example sealedampoules and vials, and stored in freeze-dried (lyophilised) state, sothat only the addition of the sterile carrier liquid, for example waterfor injection purposes, immediately before use is necessary. Injectionsolutions and suspensions prepared in accordance with the recipe can beprepared from sterile powders, granules and tablets.

It goes without saying that, in addition to the above particularlymentioned constituents, the formulations may also comprise other agentsusual in the art with respect to the particular type of formulation;thus, for example, formulations which are suitable for oraladministration may comprise flavours.

A therapeutically effective amount of a compound according to theinvention depends on a number of factors, including, for example, theage and weight of the animal, the precise condition that requirestreatment, and its severity, the nature of the formulation and themethod of administration, and is ultimately determined by the treatingdoctor or vet. However, an effective amount of a compound according tothe invention for the treatment of neoplastic growth, for example colonor breast carcinoma, is generally in the range from 0.1 to 100 mg/kg ofbody weight of the recipient (mammal) per day and particularly typicallyin the range from 1 to 10 mg/kg of body weight per day. Thus, the actualamount per day for an adult mammal weighing 70 kg is usually between 70and 700 mg, where this amount can be administered as a single dose perday or usually in a series of part-doses (such as, for example, two,three, four, five or six) per day, so that the total daily dose is thesame. An effective amount of a salt or solvate or of a physiologicallyfunctional derivative thereof can be determined as the fraction of theeffective amount of the compound according to the invention per se. Itcan be assumed that similar doses are suitable for the treatment of theother conditions mentioned above.

The invention furthermore relates to medicaments comprising at least onecompound according to the invention and/or pharmaceutically usablederivatives, solvates and stereoisomers thereof, including mixturesthereof in all ratios, and at least one further medicament activeingredient.

The invention also relates to a set (kit) consisting of separate packsof

-   -   (a) an effective amount of a compound according to the invention        and/or pharmaceutically usable derivatives, solvates and        stereoisomers thereof, including mixtures thereof in all ratios,        and    -   (b) an effective amount of a further medicament active        ingredient.

The set comprises suitable containers, such as boxes, individualbottles, bags or ampoules. The set may, for example, comprise separateampoules, each containing an effective amount of a compound according tothe invention and/or pharmaceutically usable derivatives, solvates andstereoisomers thereof, including mixtures thereof in all ratios, and aneffective amount of a further medicament active ingredient in dissolvedor lyophilised form.

Use

The compounds according to the invention and pharmaceutically usablederivatives, salts, solvates, tautomers and stereoisomers thereof,including mixtures thereof in all ratios,

are suitable as pharmaceutical active ingredients for mammals, inparticular for humans, for the preparation of a medicament for thetreatment and/or combating of cancer, tumour growth, metastatic growth,fibrosis, restenosis, HIV infection, Alzheimer's, atherosclerosis and/orfor promoting wound healing.

Particular preference is given to the use for the treatment of adisease, where the disease is a solid tumour.

The solid tumour is preferably selected from the group of tumours of thesquamous epithelium, the bladder, the stomach, the kidneys, of head andneck, the oesophagus, the cervix, the thyroid, the intestine, the liver,the brain, the prostate, the urogenital tract, the lymphatic system, thestomach, the larynx and/or the lung.

The solid tumour is furthermore preferably selected from the group lungadenocarcinoma, small-cell lung carcinomas, pancreatic cancer,glioblastomas, colon carcinoma and breast carcinoma.

Preference is furthermore given to the use for the treatment of a tumourof the blood and immune system, preferably for the treatment of a tumourselected from the group of acute myeloid leukaemia, chronic myeloidleukaemia, acute lymphatic leukaemia and/or chronic lymphatic leukaemia.

The present compounds are also suitable for combination with knownanti-cancer agents. These known anticancer agents include the following:oestrogen receptor modulators, androgen receptor modulators, retinoidreceptor modulators, cytotoxic agents, antiproliferative agents,prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors, HIVprotease inhibitors, reverse transcriptase inhibitors and furtherangiogenesis inhibitors. The present compounds are particularly suitablefor administration at the same time as radiotherapy. The synergisticeffects of inhibiting VEGF in combination with radiotherapy have beendescribed in the art (see WO 00/61186).

“Oestrogen receptor modulators” refers to compounds which interfere withor inhibit the binding of oestrogen to the receptor, regardless ofmechanism. Examples of oestrogen receptor modulators include, but arenot limited to, tamoxifen, raloxifene, idoxifene, LY353381, LY 117081,toremifene, fulvestrant,4-[7-(2,2-dimethyl-1-oxopropoxy-4-methyl-2-[4-[2-(1-piperidinyl)ethoxy]phenyl]-2H-1-benzopyran-3-yl]phenyl2,2-dimethylpropanoate,4,4′-dihydroxybenzophenone-2,4-dinitrophenylhydrazone and SH646.

“Androgen receptor modulators” refers to compounds which interfere withor inhibit the binding of androgens to the receptor, regardless ofmechanism. Examples of androgen receptor modulators include finasterideand other 5α-reductase inhibitors, nilutamide, flutamide, bicalutamide,liarozole and abiraterone acetate.

“Retinoid receptor modulators” refers to compounds which interfere withor inhibit the binding of retinoids to the receptor, regardless ofmechanism. Examples of such retinoid receptor modulators includebexarotene, tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid,α-difluoromethylornithine, ILX23-7553,trans-N-(4′-hydroxyphenyl)retinamide and N-4-carboxyphenyl-retinamide.

“Cytotoxic agents” refers to compounds which result in cell deathprimarily through direct action on the cellular function or inhibit orinterfere with cell myosis, including alkylating agents, tumour necrosisfactors, intercalators, microtubulin inhibitors and topoisomeraseinhibitors.

Examples of cytotoxic agents include, but are not limited to,tirapazimine, sertenef, cachectin, ifosfamide, tasonermin, lonidamine,carboplatin, altretamine, prednimustine, dibromodulcitol, ranimustine,fotemustine, nedaplatin, oxaliplatin, temozolomide, heptaplatin,estramustine, improsulfan tosylate, trofosfamide, nimustine,dibrospidium chloride, pumitepa, lobaplatin, satraplatin, profiromycin,cisplatin, irofulven, dexifosfamide,cis-aminedichloro(2-methylpyridine)platinum, benzylguanine,glufosfamide, GPX100,(trans,trans,trans)bis-mu-(hexane-1,6-diamine)-mu-[diamine-platinum(II)]bis[diamine(chloro)platinum(II)]tetrachloride,diarizidinylspermine, arsenic trioxide,1-(11-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine, zorubicin,idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin,pinafide, valrubicin, amrubicin, antineoplaston,3′-deamino-3′-morpholino-13-deoxo-10-hydroxycarminomycin, annamycin,galarubicin, elinafide, MEN10755 and4-demethoxy-3-deamino-3-aziridinyl-4-methylsulfonyldaunorubicin (see WO00/50032).

Examples of microtubulin inhibitors include paclitaxel, vindesinesulfate, 3′,4′-didehydro-4′-deoxy-8′-norvincaleukoblastine, docetaxol,rhizoxin, dolastatin, mivobulin isethionate, auristatin, cemadotin,RPR109881, BMS184476, vinflunine, cryptophycin,2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl)benzenesulfonamide,anhydrovinblastine,N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-t-butylamide,TDX258 and BMS188797.

Topoisomerase inhibitors are, for example, topotecan, hycaptamine,irinotecan, rubitecan,6-ethoxypropionyl-3′,4′-O-exobenzylidenechartreusin,9-methoxy-N,N-dimethyl-5-nitropyrazolo[3,4,5-kl]acridine-2-(6H)propanamine,1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3′,4′:b,7]indolizino[1,2b]quinoline-10,13(9H,15H)-dione,lurtotecan, 7-[2-(N-isopropylamino)ethyl]-(20S)camptothecin, BNP1350,BNPI1100, BN80915, BN80942, etoposide phosphate, teniposide, sobuzoxane,2′-dimethylamino-2′-deoxyetoposide, GL331,N-[2-(dimethylamino)ethyl]-9-hydroxy-5,6-dimethyl-6H-pyrido[4,3-b]carbazole-1-carboxamide,asulacrine,(5a,5aB,8aa,9b)-9-[2-[N-[2-(dimethylamino)ethyl]-N-methylamino]ethyl]-5-[4-hydroxy-3,5-dimethoxyphenyl]-5,5a,6,8,8a,9-hexohydrofuro(3′,4′:6,7)naphtho(2,3-d)-1,3-dioxol-6-one,2,3-(methylenedioxy)-5-methyl-7-hydroxy-8-methoxybenzo[c]phenanthridinium,6,9-bis[(2-aminoethyl)amino]benzo[g]isoquinoline-5,10-dione,5-(3-aminopropylamino)-7,10-dihydroxy-2-(2-hydroxyethylaminomethyl)-6H-pyrazolo[4,5,1-de]-acridin-6-one,N-[1-[2(diethylamino)ethylamino]-7-methoxy-9-oxo-9H-thioxanthen-4-ylmethyl]formamide,N-(2-(dimethylamino)ethyl)acridine-4-carboxamide,6-[[2-(dimethylamino)ethyl]amino]-3-hydroxy-7H-indeno[2,1-c]quinolin-7-oneand dimesna.

“Antiproliferative agents” include antisense RNA and DNAoligonucleotides such as G3139, ODN698, RVASKRAS, GEM231 and INX3001 andanti-metabolites such as enocitabine, carmofur, tegafur, pentostatin,doxifluridine, trimetrexate, fludarabine, capecitabine, galocitabine,cytarabine ocfosfate, fosteabine sodium hydrate, raltitrexed,paltitrexid, emitefur, tiazofurin, decitabine, nolatrexed, pemetrexed,nelzarabine, 2′-deoxy-2′-methylidenecytidine,2′-fluoromethylene-2′-deoxycytidine,N-[5-(2,3-dihydrobenzofuryl)sulfony]-N′-(3,4-dichlorophenyl)urea,N6-[4-deoxy-4-[N2-[2(E),4(E)-tetradecadienoyl]glycylamino]-L-glycero-B-L-mannohepto-pyranosyl]adenine,aplidine, ecteinascidin, troxacitabine,4-[2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimidino[5,4-b]-1,4-thiazin-6-yl-(S)-ethyl]-2,5-thienoyl-L-glutamicacid, aminopterin, 5-fluorouracil, alanosine,11-acetyl-8-(carbamoyloxymethyl)-4-formyl-6-methoxy-14-oxa-1,11-diazatetracyclo(7.4.1.0.0)tetradeca-2,4,6-trien-9-ylaceticacid ester, swainsonine, lometrexol, dexrazoxane, methioninase,2′-cyano-2′-deoxy-N4-palmitoyl-1-B-D-arabinofuransoyl cytosine and3-aminopyridine-2-carboxaldehyde thiosemicarbazone. “Antiproliferativeagents” also include monoclonal anti-bodies to growth factors other thanthose listed under “angiogenesis inhibitors”, such as trastuzumab, andtumour suppressor genes, such as p53, which can be delivered viarecombinant virus-mediated gene transfer (see U.S. Pat. No. 6,069,134,for example).

In-vitro (Enzyme) Assay for Determination of the Efficacy of theInhibitors of the Inhibition of TGF-Beta-Mediated Effects

As an example, the ability of the inhibitors to eliminateTGF-beta-mediated growth inhibition is tested.

Cells of the lung epithelial cell line Mv1Lu are sown in a defined celldensity in a 96-well microtitre plate and cultivated overnight understandard conditions. Next day, the medium is replaced by medium whichcomprises 0.5% of FCS and 1 ng/ml of TGF-beta, and the test substancesare added in defined concentrations, generally in the form of dilutionseries with 5-fold steps. The concentration of the solvent DMSO isconstant at 0.5%. After a further two days, Crystal Violet staining ofthe cells is carried out. After extraction of the Crystal Violet fromthe fixed cells, the absorption is measured spectrophotometrically at550 nm. It can be used as a quantitative measure of the adherent cellspresent and thus of the cell proliferation during the culture.

TABLE 1 Inhibition of TGF-beta Compound No. IC₅₀ [mol/l] “A1” 3.9E−06“A2” 0.2E−06 “A4” 9.9E−06 “A18” 3.3E−06 “A19” 2.5E−06

Cellular Assay for Testing TGF-Beta Receptor I Kinase Inhibitors

The kinase assay is carried out as 384-well flashplate assay. 31.2 nM ofGST-ALK5, 439 nM of GST-SMAD2 and 3 mM of ATP (with 0.3 μCi of³³P-ATP/well) are incubated in a total volume of 35 μl (20 mM of HEPES,10 mM of MgCl, 5 mM of MnCl, 1 mM of DTT, 0.1% of BSA, pH 7.4) withoutor with test substance (5-10 concentrations) at 30° C. for 45 min. Thereaction is stopped using 25 μl of 200 mM EDTA solution, filtered withsuction at room temperature after 30 min, and the wells are washed with3 times 100 μl of 0.9% NaCl solution. Radioactivity is measured in theTopCount. The IC₅₀ values are calculated using RS1.

Above and below, all temperatures are indicated in ° C. In the followingexamples, “conventional work-up” means: water is added if necessary, thepH is adjusted, if necessary, to values between 2 and 10, depending onthe constitution of the end product, the mixture is extracted with ethylacetate or dichloromethane, the phases are separated, the organic phaseis dried over sodium sulfate and evaporated, and the product is purifiedby chromatography on silica gel and/or by crystallisation. Rf values onsilica gel; eluent: ethyl acetate/methanol 9:1.

Mass spectrometry (MS): EI (electron impact ionisation) M⁺

-   -   FAB (fast atom bombardment) (M+H)³⁰    -   ESI (electrospray ionisation) (M+H)⁺

APCI-MS (atmospheric pressure chemical ionisation-mass spectrometry)(M+H)⁺.

Retention time R_(t) [min]: Determination is carried out by HPLC

Column: Chromolith Speed ROD, 50×4.6 mm² (Order No. 1.51450.0001) fromMerck

Gradient: 5.0 min, t=0 min, A:B=95:5, t=4.4 min: A:B=25:75, t=4.5 min tot=5.0 min: A:B=0:100

Flow rate: 3.00 ml/min

Eluent A: water+0.1% of TFA (trifluoroacetic acid),

Eluent B: acetonitrile+0.08% of TFA

Wavelength: 220 nm

Instead of the nitration (HNO₃/H₂SO₄), bromination is also possible withsubsequent exchange by CN or OMe (see Examples).

EXAMPLE 1

Preparation of8-methoxy-4-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenz[e]azulene(“A1”)

1.1 1.00 g of sodium are dissolved in 10 ml of methanol in a 100 mlthree-necked flask which has been rendered inert using nitrogen. Themixture is warmed to 35° C. 1.00 g of7-bromo-3-methyl-2,3,4,5-tetrahydro-1H-1-benzazepin-2-one [preparationin accordance with K. Hino et al., Chem. Pharm. Bull. 36, 2386-2400,1988] are dissolved in 12 ml of DMF. When the sodium has completelydissolved, the benzazepinone is added. 1.47 g of copper(I) iodide aresubsequently added. The reaction mixture is heated to 120° C. (oil-bathtemp.) (reflux) and stirred under these conditions overnight.

For work-up, the reaction is cooled to RT. The mixture is filteredthrough a frit covered with kieselguhr with suction. The frit is rinsedwell with DMF. The filtrate is evaporated in a high-vacuum rotaryevaporator. The residue is taken up in dichloromethane and extractedwith sodium hydroxide solution, w=2%. The aqueous phase ispost-extracted with DCM. The combined organic phases are washed withconc. sodium chloride solution, dried over sodium sulfate, filtered andevaporated to dryness.

The residue is triturated with petroleum benzine, filtered off withsuction and rinsed well.

Yield: 0.6610 g of violet crystals7-methoxy-3-methyl-2,3,4,5-tetrahydro-1H-1-benzazepin-2-one.

1.2 655 mg of7-methoxy-3-methyl-2,3,4,5-tetrahydro-1H-1-benzazepin-2-one aresuspended in 15 ml of toluene in a 100 ml round-bottomed flask. 1.012 gof 2,4-bis(4-phenoxyphenyl)-1,3,2,4-dithiadiphosphetane 2,4-disulfideare added. The yellow suspension is heated to reflux with stirring andboiled for 1.5 h.

The batch is subsequently poured into ice-water and subjected toconventional work-up.

Yield: 1.5645 g of7-methoxy-3-methyl-1,3,4,5-tetrahydro-1-benzazepine-2-thione.

1.3 1.5645 g of7-methoxy-3-methyl-1,3,4,5-tetrahydro-1-benzazepine-2-thione aredissolved in 20 ml of 1-butanol in a 100 ml round-bottomed flask. 1.68 gof 6-methylpyridine-2-carbohydrazide are added. The reaction mixture isheated to reflux and boiled at 120° C. (oil-bath temp.) overnight. Forwork-up, the batch is cooled to 40° C. with stirring and diluted with 50ml of ethyl acetate. The red-brown solution is washed three times with20 ml of semi-conc. sodium chloride solution each time. The organicphase is dried over sodium sulfate, filtered and evaporated to dryness.

The residue is taken up in ethyl acetate and extracted twice with 5%citric acid. Finally, the pH of the organic phase is restored to thebasic region by shaking with semi-conc. bicarbonate solution. Theorganic phase is dried over sodium sulfate, filtered and evaporated todryness. The residue is chromatographed on silica gel.

Column conditions:

Length: 30 cm diameter: 3.5 cm packing: silica gel 60 (0.04-0.063 mm)

Eluent: DCM/methanol, 95:5

585 mg of “A1”, EI-MS [M⁺]320, are obtained

¹H-NMR (500 MHz, DMSO-d₆) δ [ppm]: 7.8 (1H, t), 7.7 (1H,d), 7.3 (1H,d),7.0 (1H,d), 6.95 (1H,d), 6.8 (1H,dd), 3.85 (3H,s), 2.7 (3H,m), 2.4 (1H,m), 2.3 (3H,s), 1.9 (1 H,m), 1.3 (3H,br).

EXAMPLE 2

Preparation of8-hydroxy-4-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene(“A2”)

400 mg of8-methoxy-4-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azuleneare dissolved in 15 ml of dried dichloromethane in a 100 mlround-bottomed flask. 0.56 ml of boron tribromide is added. The solutionis stirred at RT for 4 hours.

The batch is then poured into a mixture of ice-water and conc.bicarbonate solution and subjected to conventional work-up.

The residue is purified by flash chromatography.

Column conditions:

Length: 30 cm, diameter: 3.5 cm, packing: silica gel 60 (0.04-0.063 mm)

Eluent: DCM/methanol, 95:5 (2 I)

305 mg of yellowish oil are obtained. The oil is purified by preparativeHPLC, giving 119 mg of “A2” as TFA salt.

Content HPLC: 99.6%; HPLC-MS [M+H⁺]307;

¹H-NMR (500 MHz, DMSO-d₆) δ [ppm]: 9.7 (1H,s), 7.8 (1H, t), 7.7 (1H,d),7.3 (1 H,d), 6.9 (1 H,d), 6.8 (1 H,d), 6.6 (1 H,dd), 2.7 (3H,m), 2.4 (1H,m), 2.3 (3H,s), 1.8 (1 H,m), 1.3 (3H,br).

EXAMPLE 3

Preparation of8-fluoro-5-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene(“A3”)

3.1 1.2884 g of 4-methyl-1,3,4,5-tetrahydro-1-benzazepin-2-one aredissolved in sulfuric acid (20 ml), and the solution is cooled to −10°C. HNO₃ (1.6 ml) is then slowly added via a dropping funnel. The mixtureis allowed to stir for a further 15 min.

The reaction solution is added to ice-water, and the precipitate formedis filtered off and washed well with water, giving 1.3852 g of4-methyl-7-nitro-1,3,4,5-tetrahydro-1-benzazepin-2-one.

This substance is dissolved in methanol and hydrogenated (Pd/C 5%),giving 770 mg of 4-methyl-7-amino-1,3,4,5-tetrahydro-1-benzazepin-2-one;¹H-NMR (500 MHz, DMSO-d₆) δ [ppm]: 9.0 (1H,s), 6.6 (1H,d), 6.41 (1H,s),6.39 (1 H,d), 4.4 (2H,br), 2.7 (2H,m), 2.4 (1 H,m), 2.1 (2H,m), 1.7 (1H,m), 1.0 (3H,d).

3.2 453 mg of nitrosyl tetrafluoroborate are dissolved in 40 ml of driedacetonitrile in a 250 ml three-necked flask with dropping funnel,thermometer and drying tube. The solution is cooled to 0° C. withstirring. The suspension of 0.76 g of4-methyl-7-amino-1,3,4,5-tetrahydro-1-benzazepin-2-one in 20 ml of driedacetonitrile is now added dropwise. The temperature is held between −2and 0° C. using the cold bath. The mixture is stirred at thistemperature for a further 45 min.

For work-up of the diazonium salt, 100 ml of dried diethyl ether isadded to the solution. The mixture is stirred for 30 min. and allowed towarm to RT. The batch solution is concentrated in a rotary evaporator.

The oily residue obtained is added in portions to 60 ml of boilingtoluene. After the addition, the mixture is boiled under reflux for afurther 15 min. The reaction mixture is cooled to RT and filteredthrough kieselguhr (Celite). The filter cake is rinsed with severalportions of chloroform. The filtrate is evaporated to dryness. The crudeproduct is purified by flash chromatography.

Column conditions:

Length: 30 cm, diameter: 3.5 cm, packing: silica gel 60 (0.04-0.063 mm)

Eluent: PE/EA, 7:3 (1 l)

144 mg of 7-fluoro-4-methyl-1,3,4,5-tetrahydro-1-benzazepin-2-one areobtained;

¹H-NMR (500 MHz, DMSO-d6) δ [ppm]: 7.1 (1H,dd), 7.05 (1H,m), 6.97(1H,dd), 2.8 (1H,m), 2.45 (1H,m), 2.3 (2H,m), 2.2 (1H,m), 1.8 (1H,m),1.0 (3H,d).

3.3 Reaction of 144 mg of7-fluoro-4-methyl-1,3,4,5-tetrahydro-1-benzazepin-2-one, analogously toExample 1, with the modified Lawesson reagent and subsequent conversioninto the triazole gives 32 mg of8-fluoro-5-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene(“A3”); EI-MS [M⁺]308;

¹H-NMR (500 MHz, DMSO-d₆) δ [ppm]: 7.8 (2H,m), 7.3 (2H,m), 7.2 (2H,m),3.1 (1 H,br), 2.8 (3H,br), 2.2 (3H,s),1.9 (1 H,br), 1.0 (3H,d).

EXAMPLE 4

Preparation of8-chloro-6-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene(“A4”)

2.9 g of 5-methyl-1,3,4,5-tetrahydro-1-benzazepin-2-one are nitratedanalogously to Example 3.1. Purification by silica-gel chromatographygives 1.9 g of 5-methyl-7-nitro-1,3,4,5-tetrahydro-1-benzazepin-2-one;

¹H-NMR (500 MHz, DMSO-d₆) δ [ppm]: 10.1 (1H, s), 8.13 (1H,dd), 8.07 (1H,d), 7.2 (1H,d), 3.1 (1H,m), 2.2 (3H,s), 1.8 (1 H,m), 1.4 (3H,d).

The substance is hydrogenated to5-methyl-7-amino-1,3,4,5-tetrahydro-1-benzazepin-2-one.

1.2 g of the amino compound are dissolved in 30 ml of hydrochloric acidand cooled to 0° C. A cold solution of sodium nitrite (439.1 mg) inwater is added dropwise at 0 to 5° C. A brown solution forms. When theaddition is complete, the mixture is stirred at about 0° C. for afurther 30 min. A cold solution of Cu(I) chloride (873.5 mg) in 20 ml ofhydrochloric acid is added dropwise at 0° C. to the diazonium solutionprepared. When the addition is complete, the mixture is allowed to warmslowly to RT. For work-up, the reaction solution is taken up in 100 mlof water and washed 3× with ethyl acetate. The combined organic phasesare washed 2× with 1N HCl and with water. The mixture is dried oversodium sulfate, and the solvent is removed in a rotary evaporator,giving 780 mg of7-chloro-5-methyl-1,3,4,5-tetrahydro-1-benzazepin-2-one.

Further reaction of 250 mg of the product with the modified Lawessonreagent and subsequent reaction with 6-methylpyridine-2-carbohydrazidegives 80 mg of8-chloro-6-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene,trifluoroacetate (“A4”); HPLC-MS [M+H⁺]325.

The following compounds are obtained analogously to the above examples

No. Name and/or structure MS “A5”8-Hydroxy-6-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro- EI-MS [M⁺]4H-2,3,10b-triazabenzo[e]azulene, trifluoroacetate 306

¹H-NMR (500 MHz, DMSO-d₆) δ [ppm]: 9.7(1H, s), 7.9(1H, t), 7.6(1H, d),7.55(1H, d), 7.1(1H, d), 6.9(1H, d), 6.7 (1H, dd), 3.2(1H, m), 2.9(1H,m), 2.5(1H, m), 2.45(3H, s), 2.4(1H, m), 1.9(1H, m), 1.3(3H, br) “A6”1-(6-Methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triaza- EI-MS [M⁺]benzo[e]azulene, trifluoroacetate 276

“A7” 8-Bromo-4-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H- EI-MS[M⁺] 2,3,10b-triazabenzo[e]azulene 369

“A8” 4-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b- EI-MS[M⁺] triazabenzo[e]azulene, bistrifluoroacetate 290 “A9”6-Methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b- HPLC-MStriazabenzo[e]azulenes [M + H⁺] 291 “A10”8-Bromo-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b- HPLC-MStriazabenzo[e]azulene, trifluoroacetate [M + H⁺] 356 “A11”8-Bromo-5-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H- HPLC-MS2,3,10b-triazabenzo[e]azulene [M + H⁺] 370 'A12”7-Methoxy-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b- EI-MS [M ⁺]triazabenzo[e]azulene 306 “A13”7-Hydroxy-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b- HPLC-MStriazabenzo[e]azulene [M + H⁺] 293 “A14”8-Cyano-4-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H- EI-MS [M⁺]2,3,10b-triazabenzo[e]azulene, trifluoroacetate 315 “A15”8-Cyano-5-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H- HPLC-MS2,3,10b-triazabenzo[e]azulene, trifluoroacetate [M + H⁺] 316 “A16”7-Acetoxy-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b- HPLC-MStriazabenzo[e]azulene [M + H⁺] 335 “A17”8-Chloro-5-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H- HPLC-MS2,3,10b-triazabenzo[e]azulene [M + H⁺] 325 “A18”8-Methoxy-6-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro- HPLC-MS4H-2,3,10b-triazabenzo[e]azulene, trifluoroacetate [M + H⁺] 321 “A19”8-Hydroxy-4-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A20”8-Hydroxy-4-propyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A21”8-Hydroxy-4-isobutyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene

The following examples relate to medicaments:

EXAMPLE A Injection vials

A solution of 100 g of an active ingredient of the formula I and 5 g ofdisodium hydrogenphosphate in 3 l of bidistilled water is adjusted to pH6.5 using 2 N hydrochloric acid, sterile filtered, transferred intoinjection vials, lyophilised under sterile conditions and sealed understerile conditions. Each injection vial contains 5 mg of activeingredient.

EXAMPLE B Suppositories

A mixture of 20 g of an active ingredient of the formula I with 100 g ofsoya lecithin and 1400 g of cocoa butter is melted, poured into mouldsand allowed to cool. Each suppository contains 20 mg of activeingredient.

EXAMPLE C Solution

A solution is prepared from 1 g of an active ingredient of the formulaI, 9.38 g of NaH₂PO₄.2 H₂O, 28.48 g of Na₂HPO₄.12 H₂O and 0.1 g ofbenzalkonium chloride in 940 ml of bidistilled water. The pH is adjustedto 6.8, and the solution is made up to 1 l and sterilised byirradiation. This solution can be used in the form of eye drops.

EXAMPLE D Ointment

500 mg of an active ingredient of the formula I are mixed with 99.5 g ofVaseline under aseptic conditions.

EXAMPLE E Tablets

A mixture of 1 kg of active ingredient of the formula I, 4 kg oflactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesiumstearate is pressed in a conventional manner to give tablets in such away that each tablet contains 10 mg of active ingredient.

EXAMPLE F Dragees

Tablets are pressed analogously to Example E and subsequently coated ina conventional manner with a coating of sucrose, potato starch, talc,tragacanth and dye.

EXAMPLE G Capsules

2 kg of active ingredient of the formula I are introduced into hardgelatine capsules in a conventional manner in such a way that eachcapsule contains 20 mg of the active ingredient.

EXAMPLE H Ampoules

A solution of 1 kg of active ingredient of the formula I in 60 l ofbidistilled water is sterile filtered, transferred into ampoules,lyophilised under sterile conditions and sealed under sterileconditions. Each ampoule contains 10 mg of active ingredient.

1. Compounds of the formula I

in which R¹ denotes H, A, OH, OA, NO₂, NH₂, NHA, NA₂, Hal, CN, A-COO,COOH, COOA or CONR⁴R⁵, R², R³ each, independently of one another, denoteH, A, alkenyl having 2-6 C atoms, alkynyl having 2-6 C atoms or Hal, R⁴,R⁵ each, independently of one another, denote H or A, A denotesunbranched or branched alkyl having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 Catoms, in which 1-7 H atoms may be replaced by F, Hal denotes F, Cl, Bror I, and pharmaceutically usable derivatives, solvates, salts,tautomers and stereoisomers thereof, including mixtures thereof in allratios.
 2. Compounds according to claim 1 in which R¹ denotes H, OH, OA,Hal, CN or A-COO, and pharmaceutically usable derivatives, solvates,salts and stereoisomers thereof, including mixtures thereof in allratios.
 3. Compounds according to claim 1 in which R² denotes H or A,and pharmaceutically usable derivatives, solvates, salts andstereoisomers thereof, including mixtures thereof in all ratios. 4.Compounds according to claim 1, in which R³ denotes A, andpharmaceutically usable derivatives, solvates, salts and stereoisomersthereof, including mixtures thereof in all ratios.
 5. Compoundsaccording to claim 1 in which A unbranched or branched alkyl having 1,2, 3, 4, 5 or 6 C atoms, in which 1-5 H atoms may be replaced by F, andpharmaceutically usable derivatives, solvates, salts and stereoisomersthereof, including mixtures thereof in all ratios.
 6. Compoundsaccording to claim 1 in which R¹ denotes H, OH, OA, Hal, CN or A-COO, R²denotes H or A, R³ denotes A, A denotes unbranched or branched alkylhaving 1, 2, 3, 4, 5 or 6 C atoms, in which 1-5 H atoms may be replacedby F, Hal denotes F, Cl, Br or I, and pharmaceutically usablederivatives, solvates, salts and stereoisomers thereof, includingmixtures thereof in all ratios.
 7. Compounds according to claim 1selected from the group No. Name and/or structure “A1”8-Methoxy-4-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A2”8-Hydroxy-4-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A3”8-Fluoro-5-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-H-2,3,10b-triazabenzo[e]azulene “A4”8-Chloro-6-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A5”8-Hydroxy-6-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene, trifluoroacetate

“A6” 1-(6-Methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triaza-benzo[e]azulene, trifluoroacetate

“A7” 8-Bromo-4-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene

“A8” 4-Methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene, bistrifluoroacetate “A9”6-Methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulenes “A10”8-Bromo-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene, trifluoroacetate “A11”8-Bromo-5-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A12”7-Methoxy-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A13”7-Hydroxy-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A14”8-Cyano-4-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene, trifluoroacetate “A15”8-Cyano-5-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene, trifluoroacetate “A16”7-Acetoxy-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A17”8-Chloro-5-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A18”8-Methoxy-6-methyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene, trifluoroacetate “A19”8-Hydroxy-4-ethyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A20”8-Hydroxy-4-propyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene “A21”8-Hydroxy-4-isobutyl-1-(6-methylpyridin-2-yl)-5,6-dihydro-4H-2,3,10b-triazabenzo[e]azulene

and pharmaceutically usable derivatives, solvates, salts andstereoisomers thereof, including mixtures thereof in all ratios. 8.Process for the preparation of compounds of the formula I according toclaim 1 and pharmaceutically usable derivatives, salts, solvates,tautomers and stereoisomers thereof, characterised in that a) a compoundof the formula II

in which R¹ and R² has the meaning indicated in claim 1, is reacted witha compound of the formula III

in which R³ has the meaning indicated in claim 1, or b) a radical R¹ isconverted into another radical R¹ by cleaving an ether, and/or a base oracid of the formula I is converted into one of its salts.
 9. Medicamentcomprising at least one compound of the formula I according to one ormore of claim 1 and/or pharmaceutically usable derivatives, solvates,salts and stereoisomers thereof, including mixtures thereof in allratios, and optionally excipients and/or adjuvants.
 10. A method ofusing compounds according to claim 1 and pharmaceutically usablederivatives, salts, solvates, tautomers and stereoisomers thereof,including mixtures thereof in all ratios, comprising preparing amedicament for the treatment and/or combating of cancer, tumour growth,metastatic growth, fibrosis, restenosis, HIV infection, Alzheimer's,atherosclerosis, and/or for promoting wound healing with a compound ofclinical pharmaceutically usable derivative, salt, solvate, tautomer,stereoisomer, or mixture thereof.
 11. A method according to claim 10,where the tumour is selected from the group of tumours of the squamousepithelium, the bladder, the stomach, the kidneys, of head and neck, theoesophagus, the cervix, the thyroid, the intestine, the liver, thebrain, the prostate, the urogenital tract, the lymphatic system, thestomach, the larynx, the lung, lung adenocarcinoma, small-cell lungcarcinoma, pancreatic cancer, glioblastoma, colon carcinoma, breastcarcinoma, tumour of the blood and immune system, acute myeloidleukaemia, chronic myeloid leukaemia, acute lymphatic leukaemia, chroniclymphatic leukaemia.
 12. A method according to claim 10 for thepreparation of a medicament for the treatment of solid tumours, where atherapeutically effective amount of a compound of the formula I isadministered in combination with a compound from the group 1) oestrogenreceptor modulator, 2) androgen receptor modulator, 3) retinoid receptormodulator, 4) cytotoxic agent, 5) antiproliferative agent, 6)prenyl-protein transferase inhibitor, 7) HMG-CoA reductase inhibitor, 8)HIV protease inhibitor, 9) reverse transcriptase inhibitor and 10)further angiogenesis inhibitor.
 13. A method according to claim 10 forthe preparation of a medicament for the treatment of solid tumours,where a therapeutically effective amount of a compound of the formula Iis administered in combination with radiotherapy and a compound from thegroup 1) oestrogen receptor modulator, 2) androgen receptor modulator,3) retinoid receptor modulator, 4) cytotoxic agent, 5) antiproliferativeagent, 6) prenyl-protein transferase inhibitor, 7) HMG-CoA reductaseinhibitor, 8) HW protease inhibitor, 9) reverse transcriptase inhibitorand 10) further angiogenesis inhibitor.
 14. Medicament comprising atleast one compound of the formula I according to one or more of claim 1and/or pharmaceutically usable derivatives, solvates and stereoisomersthereof, including mixtures thereof in all ratios, and at least onefurther medicament active ingredient.